CN114577923A - Kit for combined detection of chlorpromazine, quetiapine and dealkylated quetiapine concentration in serum and detection method - Google Patents
Kit for combined detection of chlorpromazine, quetiapine and dealkylated quetiapine concentration in serum and detection method Download PDFInfo
- Publication number
- CN114577923A CN114577923A CN202111587574.XA CN202111587574A CN114577923A CN 114577923 A CN114577923 A CN 114577923A CN 202111587574 A CN202111587574 A CN 202111587574A CN 114577923 A CN114577923 A CN 114577923A
- Authority
- CN
- China
- Prior art keywords
- quetiapine
- chlorpromazine
- dealkylated
- sample
- kit
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/30—Control of physical parameters of the fluid carrier of temperature
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/32—Control of physical parameters of the fluid carrier of pressure or speed
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
- G01N30/7233—Mass spectrometers interfaced to liquid or supercritical fluid chromatograph
- G01N30/724—Nebulising, aerosol formation or ionisation
- G01N30/726—Nebulising, aerosol formation or ionisation by electrical or glow discharge
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N2030/042—Standards
- G01N2030/045—Standards internal
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N2030/062—Preparation extracting sample from raw material
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/30—Control of physical parameters of the fluid carrier of temperature
- G01N2030/3007—Control of physical parameters of the fluid carrier of temperature same temperature for whole column
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/32—Control of physical parameters of the fluid carrier of pressure or speed
- G01N2030/324—Control of physical parameters of the fluid carrier of pressure or speed speed, flow rate
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
- G01N2030/8809—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample
- G01N2030/8813—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample biological materials
- G01N2030/8822—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample biological materials involving blood
Landscapes
- Chemical & Material Sciences (AREA)
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Dispersion Chemistry (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a kit and a detection method for jointly detecting the concentrations of chlorpromazine, quetiapine and dealkylated quetiapine in serum, wherein the kit comprises a box body, a lining, a sample extraction liquid, a calibration product, a quality control product and a sample processing board, wherein the lining is arranged at the bottom end inside the box body, a plurality of accommodating grooves are formed in the lining, and a wide-mouth bottle, a brown glass bottle and a green glass bottle are respectively arranged in the accommodating grooves; wherein, the sample extract is deposited in the inside of wide-necked bottle, and the calibrator is deposited in the inside of brown glass bottle, and the quality control article is deposited in the inside of green glass bottle. According to the detection method of the kit provided by the invention, a sample is pretreated, and then the concentrations of chlorpromazine, quetiapine and dealkylated quetiapine in serum are quantitatively detected by adopting an internal standard method and combining high performance liquid chromatography-tandem mass spectrometry. The method can carry out combined detection on the concentrations of the chlorpromazine, the quetiapine and the dealkylated quetiapine, is simple and convenient to operate, greatly reduces the detection cost and improves the detection efficiency.
Description
Technical Field
The invention relates to the field of in-vitro diagnosis, in particular to a kit and a detection method for jointly detecting the concentrations of chlorpromazine, quetiapine and dealkylated quetiapine in serum.
Background
Quetiapine is a novel phenothiazine antipsychotic, which can block a plurality of receptors such as d1 and d2 of central dopamine (dopamine) and 5-hydroxytryptamine (5-ht) 5-ht2 and 5-ht1 to play roles, is clinically effective on bipolar affective disorder and major depressive disorder (mdd), is a first-line atypical antipsychotic and a second-line antidepressant which are widely used for treating schizophrenia, has good curative effects on negative and positive symptoms and cognitive disorder of schizophrenia patients, has high treatment efficiency on refractory schizophrenia patients, has few extrapyramidal symptoms due to the fact that quetiapine is not combined with M receptors, has light anticholinergic effect and rare granulocytopenia, and is widely applied to many countries as a first-choice drug for treating schizophrenia due to the advantages of quetiapine. However, quetiapine has the function of blocking an alpha 1 adrenoceptor, and side effects such as orthostatic hypotension and dizziness can occur, so that patient monitoring is required, and therefore, the concentration detection of quetiapine is one of important indexes for patient monitoring. Since the active metabolite dealkylated quetiapine of quetiapine is also pharmacologically active and also affects the therapeutic effect, it is necessary to measure the concentration of the active metabolite dealkylated quetiapine of quetiapine.
Chlorpromazine, also known as 3- (2-chloro-10H-phenothiazin-10-yl) -N, N-dimethylpropan-1-amine, is a typical phenothiazine drug, is an antagonist of central dopamine receptors, and has various pharmacological activities. Chlorpromazine is a classical anti-schizophrenia drug, beginning to be used clinically in the 50 s of the 20 th century, and its pharmacological actions are mainly related to its dopamine (DA2) receptor blocking the limbic system and mesocortical pathways of the brain. Is mainly used for treating schizophrenia, mania, intractable hiccup and emesis clinically, and has tranquilizing, cooling and antishock effects. According to related reports, different individuals take chlorpromazine with the same dose orally, and the blood concentration can be different by more than 10 times. Therefore, the chlorpromazine 'therapeutic drug monitoring' (TDM) is developed to guide the clinical rational medication, and has positive significance.
At present, related detection methods are available at home and abroad for detecting the blood concentration of quetiapine and chlorpromazine. However, the existing detection methods have the following defects: 1. the cost of the method is too high, and the clinical popularization is limited; 2. for example, most methods for measuring the blood concentration of chlorpromazine adopt high performance liquid chromatography, but a liquid-liquid extraction method is generally adopted to treat a blood sample, so that the operation is more complicated and the method is not suitable for wide development. In addition, for the concentration detection of chlorpromazine, quetiapine and the dealkylated quetiapine which is an active metabolite of quetiapine, no detection method capable of jointly detecting the concentrations of the drugs exists in the prior art at present, and when the clinical treatment effect is judged, the detection is needed respectively, so that the efficiency is obviously lower.
In summary, there is an urgent need in the art for a detection kit and a detection method for chlorpromazine, quetiapine and dealkylated quetiapine, which have the advantages of low cost, simple sample treatment method, convenient operation, accurate result and capability of performing joint detection.
Disclosure of Invention
In order to solve the technical problems, the invention provides a kit and a detection method for combined detection of chlorpromazine, quetiapine and dealkylated quetiapine concentrations in serum, and the kit for combined detection of chlorpromazine, quetiapine and dealkylated quetiapine concentrations in serum comprises: the sample extraction device comprises a box body, a lining, a sample extraction liquid, a calibration product, a quality control product and a sample processing plate, wherein the lining is arranged at the bottom end inside the box body, a plurality of accommodating grooves are formed in the lining, and wide-mouth bottles, brown glass bottles and green glass bottles are respectively placed in the accommodating grooves;
the sample extraction liquid is stored in the wide-mouth bottle, and at least one wide-mouth bottle is arranged;
the calibrator is stored in the brown glass bottle, and at least five brown glass bottles are arranged; the quality control product is stored in the green glass bottle, and the number of the green glass bottle is at least four.
Specifically, the sample processing plate is a 96-well plate, and the 96-well plate is arranged at the bottom of the box body.
Specifically, the box body further comprises a box cover integrally formed with the box body, and the box cover is used for completely closing the top of the box body.
Specifically, the wide-mouth bottle is a 60mL wide-mouth bottle, the brown glass bottle is a 2mL brown glass bottle, and the green glass bottle is a 2mL green glass bottle.
Specifically, the color of the brown glass bottle deepens with the increase of the concentration of the calibrator, and the color of the green glass bottle deepens with the increase of the concentration of the quality control.
Specifically, still be provided with fixed inside lining in the box body, fixed inside lining is used for consolidating the wide-necked bottle.
Specifically, the sample extract is prepared by using 100% methanol, the sample extract comprises chlorpromazine-d 3, quetiapine-d 8 and dealkylated quetiapine-d 8, and the concentrations of chlorpromazine-d 3, quetiapine-d 8 and dealkylated quetiapine-d 8 are 10ng/mL, 20ng/mL and 50ng/mL respectively.
Specifically, all contain chlorpromazine, quetiapine and dealkylation quetiapine in matter accuse article and the calibrator, the matter accuse article includes high concentration matter accuse, medium concentration matter accuse, low concentration matter accuse and blank matter accuse, the concentration range of quetiapine and dealkylation quetiapine is 29 ~ 750ng/mL in the matter accuse article, the concentration range of chlorpromazine is 25 ~ 600ng/mL in the matter accuse article, the calibrator is provided with six, the concentration range of quetiapine and dealkylation quetiapine is 10 ~ 1000ng/mL in the calibrator, the concentration range of chlorpromazine is 10 ~ 800ng/mL in the matter accuse.
Specifically, the calibrator and the quality control product both contain bovine serum albumin BSA.
The invention also provides a detection method for jointly detecting the concentrations of chlorpromazine, quetiapine and dealkylated quetiapine in serum, which takes a kit for jointly detecting the concentrations of chlorpromazine, quetiapine and dealkylated quetiapine in serum as a carrier, adopts an internal standard method and combines a high performance liquid chromatography tandem mass spectrometry UPLC-MS/MS method to quantitatively detect the concentrations of chlorpromazine, quetiapine and dealkylated quetiapine in serum and comprises the following steps:
1) sample pretreatment:
firstly, after the kit and a sample to be detected are balanced to room temperature, 50 mu L of calibrator, quality control, sample to be detected and 2 blank quality control are respectively added into a sample processing plate, 300 mu L of methanol solution is added into the second blank sample, 300 mu L of sample extract is added into other samples, the mixture is oscillated and mixed at 950rpm for 10min, and then the mixture is centrifuged at 4000rpm and 4 ℃ for 10 min;
putting 100 μ L of the supernatant into a new sample processing plate, adding 200 μ L of water for redissolving, sealing the membrane, shaking and mixing at 950rpm for 5min, and centrifuging at 4000rpm and 4 ℃ for 5 min;
taking the centrifuged liquid as a sample to be analyzed, and taking 1 mu L of the centrifuged liquid for UPLC-MS/MS analysis for later use;
2) UPLC-MS/MS mass spectrometry sample injection analysis: taking 1 mu L of the sample subjected to pretreatment, and carrying out UPLC-MS/MS analysis;
3) and (4) analyzing results:
firstly, drawing a calibration curve according to the marked concentration of six calibrators, the peak area ratios of chlorpromazine, quetiapine and dealkylated quetiapine in the calibrators, and internal standard chlorpromazine-d 3, quetiapine-d 8 and dealkylated quetiapine-d 8, and fitting a calibration curve equation;
and then substituting peak area ratios of the chlorpromazine, quetiapine and dealkylated quetiapine of the detected sample and internal standards chlorpromazine-d 3, quetiapine-d 8 and dealkylated quetiapine-d 8 into a calibration curve equation, and then quantitatively calculating the concentrations of the chlorpromazine, quetiapine and dealkylated quetiapine in the sample.
Specifically, in the step 2), the detection conditions of the UPLC-MS/MS mass spectrometry are as follows:
the chromatographic column was Waters ACQUITY UPLC BEH C18, 2.1 mm. times.50 mm, 1.7 μm, the column temperature was 50 deg.C, mobile phase A was 0.1% formic acid in water, and mobile phase B was 0.1% formic acid in acetonitrile.
Specifically, in the step 2), the elution mode of the UPLC-MS/MS mass spectrometry is gradient elution, and the gradient elution procedure is as follows:
the mobile phase composition at 0min is 80% A + 20% B, the mobile phase composition at 0.2min is 80% A + 20% B, the mobile phase composition at 1.0min is 30% A + 70% B, the mobile phase composition at 1.8min is 5% A + 95% B, the mobile phase composition at 2.50min is 5% A + 95% B, the mobile phase composition at 2.51min is 80% A + 20% B, the mobile phase composition at 3.0min is 80% A + 20% B, and the flow rate in the elution process is 0.4 mL/min.
Compared with the prior art, the invention has the beneficial effects that:
1. the kit provided by the invention is reasonable in overall layout, integrates reagent materials required for detection in advance, is convenient to use and operate and carry, does not need extra dilution preparation operation for a calibrator and a quality control material, can be directly used for detecting the concentrations of chlorpromazine, quetiapine and dealkylated quetiapine in a human serum sample by liquid chromatography-mass spectrometry after sampling is subjected to conventional pretreatment operation, effectively shortens the detection time, is beneficial to improving the analysis and detection efficiency, and is strong in convenience;
2. the kit disclosed by the invention is reasonable in layout and simple in structure, is convenient for a laboratory worker to operate and carry, and improves the analysis and detection efficiency;
3. in the sample materials of the kit, the quality control substances are chlorpromazine, quetiapine and dealkylated quetiapine quality control substances, the kit is attached with three quality control substances with different concentrations of high, medium and low, simultaneously carries out quality control on the detection results of chlorpromazine, quetiapine and dealkylated quetiapine, is attached with a blank control quality control substance for specificity evaluation and monitoring of high-concentration sample residues, can be used for detection and quality control of mass samples or long-time detection in liquid chromatography-mass spectrometry, and is very convenient to use;
4 the MRM method adopted by the kit is accurate and quantitative according to the charge-to-mass ratio of the detected object, has high sensitivity and strong specificity, is not interfered by metabolites and in vivo monoclonal antibodies, and is suitable for clinical routine detection. The chlorpromazine peak time is about 0.81min, the quetiapine peak time is about 0.74min, and the dealkylation quetiapine peak time is about 0.72min, and the detection of one sample can be completed in 3min on average, so that the detection efficiency is greatly improved, and the cost is conveniently reduced.
Drawings
FIG. 1 is a schematic perspective view of one embodiment of the kit of the present invention;
FIG. 2 is a schematic structural view of the inner liner of the kit of the present invention;
FIG. 3 is a chromatogram of chlorpromazine in the detection method of the present invention;
FIG. 4 is a chromatogram of quetiapine in the detection method of the present invention;
FIG. 5 is a chromatogram of dealkylquetiapine in the detection method of the present invention;
FIG. 6 is a mass spectrum of chlorpromazine in the detection method of the invention;
FIG. 7 is a mass spectrum of quetiapine in the detection method of the present invention;
FIG. 8 is a mass spectrum of dealkylquetiapine in the detection method of the present invention;
in the figure: 1. a box body; 2. a liner; 3. a jar; 4. a brown glass bottle; 5. a green glass bottle; 6. a sample processing plate; 7. and (7) a box cover.
Detailed Description
The technical solutions of the present invention will be described clearly and completely below, and it should be apparent that the described embodiments are some, but not all, embodiments of the present invention. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention. Materials, instruments, reagents and the like used in the following examples are commercially available unless otherwise specified. The technical means used in the examples are conventional means well known to those skilled in the art, unless otherwise specified.
Example one
As shown in fig. 1-2, a kit for combined detection of chlorpromazine, quetiapine, and dealkylated quetiapine concentrations in serum, comprising: the device comprises a box body 1, a lining 2, a sample extraction liquid, a calibration product, a quality control product and a sample processing plate 6, wherein the lining 2 is arranged at the bottom end inside the box body 1, a plurality of accommodating grooves are formed in the lining 2, and a wide-mouth bottle 3, a brown glass bottle 4 and a green glass bottle 5 are respectively arranged in the accommodating grooves;
wherein, the sample extract is stored in the wide-mouth bottle 3, and at least one wide-mouth bottle 3 is arranged;
the calibrator is stored in the brown glass bottle 4, and at least five brown glass bottles 4 are arranged; the quality control products are stored in the green glass bottles 5, and at least four green glass bottles 5 are arranged; the sample processing plate 6 is a 96-hole plate, and the 96-hole plate is arranged at the bottom of the box body 1; the box body 1 also comprises a box cover 7 which is integrally formed with the box body 1, and the box cover 7 is used for completely sealing the top of the box body 1; the wide-mouth bottle 3 is a 60mL wide-mouth bottle 3, the brown glass bottle 4 is a 2mL brown glass bottle, and the green glass bottle 5 is a 2mL green glass bottle; the color of the brown glass bottle 4 is deepened along with the increase of the concentration of the calibrator, and the color of the green glass bottle 5 is deepened along with the increase of the concentration of the quality control material; still be provided with fixed inside lining in the box body 1, fixed inside lining is used for consolidating wide-necked bottle 3, and fixed inside lining adopts the hermite material, only is equipped with a storage tank on the fixed inside lining because the height and the sample extraction liquid sample bottle height difference of standard article and matter accuse article sample bottle to this further fixed not hard up that prevents the sample bottle.
The whole layout of the kit is reasonable, reagents and consumables required in the whole detection process are integrated in advance, the operation of a laboratory technician is facilitated, the kit is convenient to carry, the time for pre-preparing the reagents in advance is effectively saved, meanwhile, the detection error caused by reagent preparation is avoided, the detection time consumption is effectively shortened, and the detection efficiency is improved.
The reagent box shown in figure 1 further comprises a box cover 7, and the box cover 7 is matched with the box body 1 to realize the function of closing or opening the reagent box. In fig. 1, the box cover 7 is connected with the box body 1. In other modified modes, the box cover 7 and the box body 1 can be independent. When the box cover 7 is matched and closed with the box body 1, the reagent box can keep a sealed state. Preferably, a protection pad made of an elastic buffer packing material is arranged on the inner side surface of the box cover 7. The protection pad can reduce the extrusion force in the vertical direction and protect the container from being damaged.
The kit provided by the invention further comprises an instruction and a target value table, wherein the instruction is recorded with composition information of the kit and a using method.
Example two kit composition and preparation method
Based on a kit of the first embodiment, the specific components and specific concentrations of the kit of this embodiment are shown in the following table:
preparation of 5% BSA solution
The actual dosage amount can be adjusted according to the actual production amount.
Weighing 250g of bovine serum albumin by using a balance, transferring the bovine serum albumin into a beaker filled with 5L of 1 multiplied by PBS, and carrying out ultrasonic treatment while stirring until the bovine serum albumin is completely dissolved; then filtering with two layers of filter paper, and then filtering with 0.22 μm water phase filter membrane; finally, NaN3 and EDTA were added to make a 5% BSA solution containing 0.09% NaN3 and 10 mg/mLEDTA. After the preparation is finished, the mixture is stored in a refrigerator at the temperature of 2-8 ℃.
Preparation of (II) Standard solution
Preparing chlorpromazine, quetiapine and dealkylated quetiapine stock solutions:
weighing 11.15mg of chlorpromazine hydrochloride (TCI) by a hundred thousand grade balance, placing the weighed chlorpromazine hydrochloride (TCI) in a 10mL volumetric flask, adding a proper amount of methanol to dissolve and dilute the chlorpromazine hydrochloride (TCI) to a scale, and reversing and uniformly mixing to obtain 1mg/mL chlorpromazine TCI stock solution 1. And (4) converting the actual concentration of the stock solution according to the actual weighed mass. Subpackaging after the preparation is finished, and storing in a refrigerator at-20 ℃.
Weighing 13.589mg of quetiapine (TCI) by a hundred thousand grade balance, placing the quetiapine (TCI) in a 10mL volumetric flask, adding a proper amount of methanol to dissolve and dilute the quetiapine (TCI) to scale, and reversing and mixing the quetiapine (TCI) and the quetiapine to obtain a quetiapine TCI stock solution 2 with the concentration of 1 mg/mL. And converting the actual concentration of the stock solution according to the actual weighed mass. Subpackaging after the preparation is finished, and storing in a refrigerator at-20 ℃.
Weighing 12.99mg of dealkylquetiapine (TCI) by a hundred thousand grade balance, placing the dealkylquetiapine (TCI) in a 10mL volumetric flask, adding a proper amount of methanol to dissolve and dilute the dealkylquetiapine (TCI) to scale, and reversing and mixing the solution uniformly to obtain a dealkylquetiapine TCI stock solution 3 with the concentration of 1 mg/mL. And converting the actual concentration of the stock solution according to the actual weighed mass. Subpackaging after the preparation is finished, and storing in a refrigerator at-20 ℃.
Preparation of (tri) chlorpromazine-d 3, quetiapine-d 8 and dealkylated quetiapine-d 8 stock solutions
Quetiapine-d 8 and dealkylated quetiapine-d 8 were placed in 10mL measuring flasks (V) at about 1mg (m) using a hundred thousand scale balance, dissolved in methanol and diluted to the scale to prepare quetiapine-d 8 and dealkylated quetiapine-d 8 internal standard stock solutions at a concentration C ═ m × purity × d8 ratio/V. The chlorpromazine-d 3 stock solution is a standard stock solution with the concentration of 100 mu g/mL, and can be directly used. The prepared product is stored in a refrigerator at the temperature of-20 ℃.
(IV) preparation of sample extract
Taking appropriate amount of chlorpromazine-d 3, quetiapine-d 8 and dealkylated quetiapine-d 8 stock solutions respectively, adding methanol, and mixing uniformly to prepare sample extract with concentration of about 10ng/mL/20ng/mL/50 ng/mL. The actual dosage amount can be adjusted according to the actual production amount. Subpackaging after the preparation is finished, and storing in a refrigerator at 4 ℃.
In this embodiment, the calibration product is provided with six calibration products, which are respectively labeled as C1, C2, C3, C4, C5, and C6, the quality control products are divided into a high-concentration quality control product HQC, a medium-concentration quality control product MQC, a low-concentration quality control product LQC, and a blank quality control product, the blank quality control product is 5% bovine serum albumin prepared in advance, and the preparation of the C1, C2, C3, C4, C5, C6, the high-concentration quality control product HQC, the medium-concentration quality control product MQC, and the low-concentration quality control product LQC is obtained by further preparing a working solution, and the preparation of the working solution is shown in the following table:
the formulations of the calibrator and quality control were as follows:
EXAMPLE III method for detecting the concentration of chlorpromazine, quetiapine and dealkylated quetiapine in serum by using the kit
The kit for jointly detecting the concentrations of chlorpromazine, quetiapine and dealkylated quetiapine in serum is used as a carrier, an internal standard method is adopted, and a high performance liquid chromatography tandem mass spectrometry UPLC-MS/MS method is combined to quantitatively detect the concentrations of chlorpromazine, quetiapine and dealkylated quetiapine in serum, and the method comprises the following steps:
1) sample pretreatment:
firstly, after the kit and a sample to be detected are balanced to room temperature, 50 mu L of calibrator, quality control, sample to be detected and 2 blank quality control are respectively added into a sample processing plate, 300 mu L of methanol solution is added into the second blank sample, 300 mu L of sample extract is added into other samples, the mixture is oscillated and mixed at 950rpm for 10min, and then the mixture is centrifuged at 4000rpm and 4 ℃ for 10 min;
putting 100 μ L of the supernatant into a new sample processing plate, adding 200 μ L of water for redissolving, sealing the membrane, shaking and mixing at 950rpm for 5min, and centrifuging at 4000rpm and 4 ℃ for 5 min;
taking 1 mu L of the centrifuged liquid as a sample to be analyzed, and using the liquid for UPLC-MS/MS analysis for later use;
2) UPLC-MS/MS mass spectrometry sample injection analysis: taking 1 mu L of the sample subjected to pretreatment, and carrying out UPLC-MS/MS analysis;
3) and (4) analyzing results:
firstly, drawing a calibration curve according to the marked concentration of six calibrators, the peak area ratios of chlorpromazine, quetiapine and dealkylated quetiapine in the calibrators, and internal standard chlorpromazine-d 3, quetiapine-d 8 and dealkylated quetiapine-d 8, and fitting a calibration curve equation;
and then substituting peak area ratios of the chlorpromazine, quetiapine and dealkylated quetiapine of the detected sample and internal standards chlorpromazine-d 3, quetiapine-d 8 and dealkylated quetiapine-d 8 into a calibration curve equation, and then quantitatively calculating the concentrations of the chlorpromazine, quetiapine and dealkylated quetiapine in the sample.
In the step 2), the detection conditions of the UPLC-MS/MS mass spectrometry are as follows:
the chromatographic column was Waters ACQUITY UPLC BEH C18, 2.1mm X50 mm, 1.7 μm, column temperature 50 deg.C, mobile phase A was 0.1% formic acid in water, and mobile phase B was 0.1% formic acid in acetonitrile.
In the step 2), the elution mode of the UPLC-MS/MS mass spectrometry is gradient elution, and the gradient elution procedure is as follows:
the mobile phase composition at 0min is 80% A + 20% B, the mobile phase composition at 0.2min is 80% A + 20% B, the mobile phase composition at 1.0min is 30% A + 70% B, the mobile phase composition at 1.8min is 5% A + 95% B, the mobile phase composition at 2.50min is 5% A + 95% B, the mobile phase composition at 2.51min is 80% A + 20% B, the mobile phase composition at 3.0min is 80% A + 20% B, and the flow rate in the elution process is 0.4 mL/min.
In one embodiment, the mass spectrometry conditions are as shown in the following table:
parameters of ion source
The MRM information is shown in the following table:
is a quantitative ion pair
As shown in fig. 3-8, in order to detect chlorpromazine, quetiapine and dealkylated quetiapine in serum by using the kit and the detection method provided by the invention, fig. 3-5 are chromatograms of chlorpromazine, quetiapine and dealkylated quetiapine, respectively, and fig. 6-8 are mass spectrograms of chlorpromazine, quetiapine and dealkylated quetiapine, respectively.
The linear experiment of the detection method provided by the invention is as follows:
after the kit and the sample to be detected are balanced to room temperature, 50 mu L of calibrator, quality control, sample to be detected and 2 blank quality control are respectively added into a 96-hole plate, 300 mu L of methanol solution is added into the second blank sample, 300 mu L of sample extract is added into other samples, the samples are oscillated and mixed at 950rpm for 10min, and then the samples are centrifuged at 4000rpm and 4 ℃ for 10 min; putting 100 μ L of the supernatant into a new 96-well plate, adding 200 μ L of water for redissolving, sealing the membrane, shaking and mixing at 950rpm for 5min, and centrifuging at 4000rpm and 4C for 5 min; and centrifuging to prepare a sample to be analyzed for blood concentration analysis of chlorpromazine, quetiapine and dealkylated quetiapine.
The chlorpromazine quantitative detection range is 10ng/mL-800ng/mL, and the quetiapine and dealkylated quetiapine quantitative detection range is 10ng/mL-1000 ng/mL. The quantitative detection range of chlorpromazine, quetiapine and dealkylated quetiapine is determined to be linear or not according to the theoretical values and the measured values of three groups of samples covering the linear range of 5% BSA (containing 0.09% NaN3 and 10 mg/mLEDTA). The measurement was performed once a day in three batches. The linear fitting correlation coefficient R is greater than 0.99, the calculated concentration of the corrected standard sample should be within +/-15% of the marked value, and the lower limit of the quantification should be within +/-20%.
Performing linear regression on the sample concentration by comparing the peak area of the sample with the peak area of the internal standard, and obtaining a linear equation as follows: chlorpromazine 0.00822157 × x +0.00782859 (R0.999294); quetiapine: 0.00897162 × x +0.0167295 (R0.999683); quetiapine: 0.00346476 × x +0.00706051 (R0.999250).
And (4) experimental conclusion: the results show that the chlorpromazine, the quetiapine and the dealkylated quetiapine have good linear correlation, R is greater than 0.99, the bias is less than +/-10 percent, and the requirements are met.
Examples stability experiments for Tetrachloropropazine, quetiapine and dealkylated quetiapine
And preparing sufficient LQC, MQC and HQC samples according to the preparation method provided by the embodiment II for later use, researching the storage conditions of the samples, respectively storing the samples in a sealed mode at the temperature of 2-8 ℃ and the temperature of 25 ℃ and the temperature of-20 ℃ and repeatedly freezing and thawing, and detecting the storage stability of the samples within a certain time.
1) Storage stability of sample at 2-8 DEG C
And (4) investigating the stability of the compound when the compound is placed at the temperature of 2-8 ℃. And preparing LQC and HQC samples by using a human-derived matrix, standing for 0, 3 and 7 days at the temperature of 2-8 ℃, and preparing 2 stability quality control samples at each concentration and time point. The stability sample detection concentration should be within ± 15% compared to the T0 sample at each time point.
2) Storage stability of samples under 25 ℃ constant temperature and humidity box conditions
The stability of the compound when left at room temperature was examined. LQC and HQC samples were prepared with human and alternative matrices, respectively, and left at room temperature (25 ℃) for 0, 4, 24, and 48h, with 3 stability quality control samples prepared at each time point for each concentration. The stability sample detection concentration should be within ± 15% of the T0 sample at each time point.
3) Storage stability of samples at-20 deg.C
The stability of the compound when placed at-20 ℃ was examined. LQC and HQC samples were prepared using human-derived matrices and placed at-20 ℃ for 0, 60 days, and 2 stability control samples were prepared at each concentration and time point. The stability sample detection concentration should be within ± 15% of the theoretical sample at each time point.
4) Sample freeze thaw stability
The compounds were examined for their freeze-thaw stability when placed at-20 ℃. Preparing LQC, MQC and HQC samples by using human-derived matrixes and alternative matrixes, storing the LQC, MQC and HQC samples at-20 ℃, taking out the LQC, MQC and HQC samples after the LQC, MQC and HQC samples are placed for at least 12h for freezing, placing the LQC, MQC and HQC samples at room temperature for thawing, taking the LQC, MQC and HQC samples as 1-time freeze-thaw experiment, respectively examining the influence on the sample concentration after 1-time, 2-time and 3-time freeze-thaw, and preparing 3 stability quality control samples at each freeze-thaw time point. The detection concentration of the stability sample after 1, 2 and 3 times of freeze thawing is within +/-15% compared with that of a sample without freeze thawing.
And (4) experimental conclusion: samples stored at 2-8 ℃ were tested for high and low values on days 0, 3, and 7, and had a bias of less than 15.0% relative to T0 (chlorpromazine low value sample was 16.13% on day three, but 11.14% on day seven, indicating that the data was more biased due to handling errors) to meet acceptable standards. The chlorpromazine, quetiapine and dealkylated quetiapine in the sample have good stability when stored for 0-7 days at the temperature of 2-8 ℃.
The sample preserved in the environment of 25 ℃ is detected at 0, 4, 24 and 48 hours and the low, medium and high value samples, the detection bias is less than 15.0 percent relative to the T0, the samples meet the acceptable standard, and the stability of the chlorpromazine, quetiapine and dealkylated quetiapine in the sample is good when the samples are preserved for 0 to 48 hours at the environment of 25 ℃.
Placing the sample at-20 ℃ for 0 and 60 days, preparing 2 stability quality control samples at each concentration and each time point, wherein the deviation of the detection concentration of the stability sample at each time point compared with the theoretical sample is within +/-15%.
Freeze-thaw stability of the samples when placed at-20 ℃. Compared with a sample which is not frozen and thawed, the stability sample after being frozen and thawed for 1 time, 2 times and 3 times has the deviation within +/-15 percent, meets the acceptable standard, and shows that the stability of the sample is good after the chlorpromazine, the quetiapine and the dealkylated quetiapine in the sample are frozen and thawed for 3 times at the temperature of-20 ℃.
In conclusion, the kit for jointly detecting the concentrations of chlorpromazine, quetiapine and dealkylated quetiapine in serum and the detection method thereof provided by the invention have the advantages of accurate quantification, good stability, capability of simultaneously completing the detection of the concentrations of chlorpromazine, quetiapine and dealkylated quetiapine in serum, great improvement on detection efficiency and reduction in detection cost.
In summary, the above embodiments are merely preferred embodiments of the present invention, and are not intended to limit the scope of the present invention, and any modifications, equivalents, improvements, etc. made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (12)
1. A kit for jointly detecting the concentrations of chlorpromazine, quetiapine and dealkylated quetiapine in serum is characterized by comprising: the device comprises a box body (1), a lining (2), a sample extraction liquid, a calibration product, a quality control product and a sample processing plate (6), wherein the lining (2) is arranged at the bottom end inside the box body (1), a plurality of accommodating grooves are formed in the lining (2), and a wide-mouth bottle (3), a brown glass bottle (4) and a green glass bottle (5) are respectively placed in the accommodating grooves;
wherein the sample extraction liquid is stored in the wide-mouth bottle (3), and at least one wide-mouth bottle (3) is arranged; the calibrator is stored inside the brown glass bottle (4), and at least five brown glass bottles (4) are arranged; the quality control product is stored in the green glass bottle (5), and the number of the green glass bottle (5) is at least four.
2. The kit for jointly detecting the concentration of chlorpromazine, quetiapine and dealkylated quetiapine in serum according to claim 1, wherein the sample processing plate (6) is a 96-well plate, and the 96-well plate is arranged at the bottom of the box body (1).
3. The kit for jointly detecting the concentrations of chlorpromazine, quetiapine and dealkylated quetiapine in serum according to claim 1, wherein the box body (1) further comprises a box cover (7) integrally formed with the box body (1), and the box cover (7) is used for completely closing the top of the box body (1).
4. The kit for jointly detecting the concentration of chlorpromazine, quetiapine and dealkylated quetiapine in serum according to claim 1, wherein the jar (3) is a 60mL jar (3), the brown glass bottle (4) is a 2mL brown glass bottle, and the green glass bottle (5) is a 2mL green glass bottle.
5. The kit for jointly detecting chlorpromazine, quetiapine and dealkylated quetiapine concentrations in serum according to claim 1, characterized in that the color of the brown glass bottle (4) deepens as the concentration of a calibrator increases, and the color of the green glass bottle (5) deepens as the concentration of a quality control increases.
6. The kit for jointly detecting the concentration of chlorpromazine, quetiapine and dealkylated quetiapine in serum according to claim 1, wherein a fixed lining is further arranged in the box body (1), and the fixed lining is used for reinforcing the wide-mouth bottle (3).
7. The kit for jointly detecting chlorpromazine, quetiapine and dealkylated quetiapine concentrations in serum according to claim 1, wherein the sample extract is prepared by using 100% methanol, the sample extract contains chlorpromazine-d 3, quetiapine-d 8 and dealkylated quetiapine-d 8, and the concentration of chlorpromazine-d 3, quetiapine-d 8 and dealkylated quetiapine-d 8 is 10ng/mL, 20ng/mL and 50ng/mL respectively.
8. The kit for jointly detecting the concentrations of chlorpromazine, quetiapine and dealkylated quetiapine in serum according to claim 1, wherein the quality control product and the calibrator both contain chlorpromazine, quetiapine and dealkylated quetiapine, the quality control product comprises a high-concentration quality control product, a medium-concentration quality control product, a low-concentration quality control product and a blank quality control product, the concentration ranges of the quetiapine and the dealkylated quetiapine in the quality control product are 29-750 ng/mL, the concentration range of the chlorpromazine in the quality control product is 25-600 ng/mL, the calibrator is provided with six components, the concentration ranges of the quetiapine and the dealkylated quetiapine in the calibrator are 10-1000 ng/mL, and the concentration range of the chlorpromazine in the quality control product is 10-800 ng/mL.
9. The kit for jointly detecting the concentrations of chlorpromazine, quetiapine and dealkylated quetiapine in serum according to claim 1, wherein the calibrator and the quality control material both further comprise bovine serum albumin BSA.
10. The detection method of the kit for jointly detecting the concentrations of chlorpromazine, quetiapine and dealkylated quetiapine in serum according to any one of claims 1 to 9, which is characterized in that the kit for jointly detecting the concentrations of chlorpromazine, quetiapine and dealkylated quetiapine in serum is used as a carrier, an internal standard method is adopted, and a high performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method is combined to quantitatively detect the concentrations of chlorpromazine, quetiapine and dealkylated quetiapine in serum, and comprises the following steps:
1) sample pretreatment:
firstly, after the kit and a sample to be detected are balanced to room temperature, 50 mu L of calibrator, quality control, sample to be detected and 2 blank quality control are respectively added into a sample processing plate, 300 mu L of methanol solution is added into the second blank sample, 300 mu L of sample extract is added into other samples, the mixture is oscillated and mixed at 950rpm for 10min, and then the mixture is centrifuged at 4000rpm and 4 ℃ for 10 min;
putting 100 μ L of the supernatant into a new sample processing plate, adding 200 μ L of water for redissolving, sealing the membrane, shaking and mixing at 950rpm for 5min, and centrifuging at 4000rpm and 4 ℃ for 5 min;
taking 1 mu L of the centrifuged liquid as a sample to be analyzed, and using the liquid for UPLC-MS/MS analysis for later use;
2) UPLC-MS/MS mass spectrum sample injection analysis: taking 1 mu L of the sample subjected to pretreatment, and carrying out UPLC-MS/MS analysis;
3) and (4) analyzing results:
firstly, drawing a calibration curve according to the marked concentration of six calibrators, the peak area ratios of chlorpromazine, quetiapine and dealkylated quetiapine in the calibrators, and internal standard chlorpromazine-d 3, quetiapine-d 8 and dealkylated quetiapine-d 8, and fitting a calibration curve equation;
and then substituting peak area ratios of the chlorpromazine, quetiapine and dealkylated quetiapine of the detected sample and internal standards chlorpromazine-d 3, quetiapine-d 8 and dealkylated quetiapine-d 8 into a calibration curve equation, and then quantitatively calculating the concentrations of the chlorpromazine, quetiapine and dealkylated quetiapine in the sample.
11. The detection method of the kit for jointly detecting the concentrations of chlorpromazine, quetiapine and dealkylated quetiapine in serum according to claim 10, wherein in the step 2), the detection conditions of the UPLC-MS/MS mass spectrometry are as follows:
the chromatographic column was Waters ACQUITY UPLC BEH C18, 2.1 mm. times.50 mm, 1.7 μm, the column temperature was 50 deg.C, mobile phase A was 0.1% formic acid in water, and mobile phase B was 0.1% formic acid in acetonitrile.
12. The detection method of the kit for jointly detecting the concentrations of chlorpromazine, quetiapine and dealkylated quetiapine in serum according to claim 10, wherein in the step 2), the elution mode of the UPLC-MS/MS mass spectrometry is gradient elution, and the gradient elution program is as follows:
the mobile phase composition at 0min is 80% A + 20% B, the mobile phase composition at 0.2min is 80% A + 20% B, the mobile phase composition at 1.0min is 30% A + 70% B, the mobile phase composition at 1.8min is 5% A + 95% B, the mobile phase composition at 2.50min is 5% A + 95% B, the mobile phase composition at 2.51min is 80% A + 20% B, the mobile phase composition at 3.0min is 80% A + 20% B, and the flow rate in the elution process is 0.4 mL/min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111587574.XA CN114577923A (en) | 2021-12-23 | 2021-12-23 | Kit for combined detection of chlorpromazine, quetiapine and dealkylated quetiapine concentration in serum and detection method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111587574.XA CN114577923A (en) | 2021-12-23 | 2021-12-23 | Kit for combined detection of chlorpromazine, quetiapine and dealkylated quetiapine concentration in serum and detection method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114577923A true CN114577923A (en) | 2022-06-03 |
Family
ID=81768863
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111587574.XA Pending CN114577923A (en) | 2021-12-23 | 2021-12-23 | Kit for combined detection of chlorpromazine, quetiapine and dealkylated quetiapine concentration in serum and detection method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114577923A (en) |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| SU726471A1 (en) * | 1977-07-18 | 1980-04-05 | Тюменский государственный медицинский институт | Method of quantitative determining of phenothiazine derivatives |
| US5413911A (en) * | 1990-12-14 | 1995-05-09 | Abbott Laboratories | Determination of tricyclic antidepressant drugs in the presence of interfering substances |
| CN109085263A (en) * | 2018-08-03 | 2018-12-25 | 杭州佰勤医疗器械有限公司 | Liquid chromatography tandem mass spectrometry detects the kit of anti-schizophrenia drug and its application in serum plasma |
| CN109668979A (en) * | 2018-12-21 | 2019-04-23 | 山东英盛生物技术有限公司 | Method that is a kind of while detecting 17 kinds of antipsychotics in blood sample |
| CN110763799A (en) * | 2019-11-29 | 2020-02-07 | 吉林和合医学检验有限公司 | Method for simultaneously detecting quetiapine and N-dealkylated quetiapine contents in blood |
| CN111239282A (en) * | 2020-02-08 | 2020-06-05 | 宁波市疾病预防控制中心 | Method and kit for determining phenobarbital in blood and application |
| CN111257486A (en) * | 2019-12-31 | 2020-06-09 | 北京回龙观医院(北京心理危机研究与干预中心) | Method and kit for detecting 5 psychotropic drugs and main metabolites in blood |
-
2021
- 2021-12-23 CN CN202111587574.XA patent/CN114577923A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| SU726471A1 (en) * | 1977-07-18 | 1980-04-05 | Тюменский государственный медицинский институт | Method of quantitative determining of phenothiazine derivatives |
| US5413911A (en) * | 1990-12-14 | 1995-05-09 | Abbott Laboratories | Determination of tricyclic antidepressant drugs in the presence of interfering substances |
| CN109085263A (en) * | 2018-08-03 | 2018-12-25 | 杭州佰勤医疗器械有限公司 | Liquid chromatography tandem mass spectrometry detects the kit of anti-schizophrenia drug and its application in serum plasma |
| CN109668979A (en) * | 2018-12-21 | 2019-04-23 | 山东英盛生物技术有限公司 | Method that is a kind of while detecting 17 kinds of antipsychotics in blood sample |
| CN110763799A (en) * | 2019-11-29 | 2020-02-07 | 吉林和合医学检验有限公司 | Method for simultaneously detecting quetiapine and N-dealkylated quetiapine contents in blood |
| CN111257486A (en) * | 2019-12-31 | 2020-06-09 | 北京回龙观医院(北京心理危机研究与干预中心) | Method and kit for detecting 5 psychotropic drugs and main metabolites in blood |
| CN111239282A (en) * | 2020-02-08 | 2020-06-05 | 宁波市疾病预防控制中心 | Method and kit for determining phenobarbital in blood and application |
Non-Patent Citations (1)
| Title |
|---|
| 温预关;廖日房;曾转萍;张明;: "HPLC-电喷雾串联质谱法快速测定人血浆中喹硫平的浓度", 中国药房, vol. 32, no. 32, pages 2504 - 2506 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Bozik et al. | Safety, tolerability, and pharmacokinetics of KNS‐760704 (dexpramipexole) in healthy adult subjects | |
| CN112946107B (en) | Analysis method of N-nitrosodimethylamine and N-nitrosodiethylamine in argatroban bulk drug or preparation | |
| CN110068644B (en) | Method for determining concentration of olanzapine in plasma by high performance liquid chromatography tandem mass spectrometry | |
| CN111766312A (en) | Method for detecting antifungal drugs in serum by ultra-high performance liquid chromatography tandem mass spectrometry technology | |
| CN115453012B (en) | Reversed-phase HPLC method for simultaneously measuring multiple positional isomers in voathixetine hydrobromide | |
| CN111707764A (en) | Detection method of cyclobenzaprine hydrochloride sustained-release capsule | |
| CN114577923A (en) | Kit for combined detection of chlorpromazine, quetiapine and dealkylated quetiapine concentration in serum and detection method | |
| CN107228909B (en) | Method for measuring acotiamide hydrochloride raw material medicine and related substances in preparation thereof by using HPLC | |
| CN114354789B (en) | Method for simultaneously measuring cabozantinib analogue and related substances thereof | |
| CN114544796B (en) | Method for measuring settop alcohol in plasma by liquid phase mass spectrometry | |
| CN114689737B (en) | Analysis method of S-o-chlorophenylglycine methyl tartrate related substances | |
| CN113514588B (en) | High performance liquid chromatography analysis method of relevant substances of cysteamine for injection | |
| El-Masry et al. | Microdetermination of hyoscyamine and scopolamine in mixtures | |
| CN114264765B (en) | Analytical method for determining related substances in glimepiride intermediate by utilizing HPLC | |
| CN107884496B (en) | Method for determining content of succinic acid in trelagliptin succinate | |
| CN113514589A (en) | High performance liquid chromatography analysis method of stannous glucoheptonate related substance for injection | |
| CN109298081B (en) | Method for determining impurity A biological sample in Cetilistat | |
| CN114113402B (en) | Method for measuring pinanediol content in bortezomib by adopting high performance liquid chromatography | |
| CN114518423B (en) | Method for detecting impurities in dopamine hydrochloride injection | |
| CN110412164A (en) | A kind of detection method of the related substance of mexiletine hydrochloride | |
| CN112611822B (en) | Detection method and application of cefoperazone sodium and sulbactam sodium related substances | |
| CN115236252B (en) | Method for detecting 25-hydroxy vitamin D in serum | |
| CN117890496B (en) | Method for detecting related substances of compound preparation of novel oral solution of guaifenesin | |
| Soore et al. | Investigation Study of Cephalexin Capsule Content from Different Drug Companies Available in the Iraqi Pharmaceutical Market | |
| CN117871727A (en) | Method for detecting citrate content in linezolid glucose injection |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |